Team:Paris Bettencourt/Notebook/Drug Screening/Tuesday 11th June.html

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<a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a>
<a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a>
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<h3>Day Number<sup>suffix</sup> Month</h3>
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<h3>Tuesday 11<sup>th</sup> June</h3>
<p><b><em>
<p><b><em>
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<br>Primers: <br>MtbfdrA F &nbsp;&nbsp;&nbsp;&nbsp; TATGACTCATATGCGTCCCTATTACATCGCCATCG
<br>Primers: <br>MtbfdrA F &nbsp;&nbsp;&nbsp;&nbsp; TATGACTCATATGCGTCCCTATTACATCGCCATCG
<br>MtbfdrA R &nbsp;&nbsp;&nbsp;&nbsp; ATCCGGGATCCTCAGCCGAGCCCAATCCGCAACAGC
<br>MtbfdrA R &nbsp;&nbsp;&nbsp;&nbsp; ATCCGGGATCCTCAGCCGAGCCCAATCCGCAACAGC
-
<br>The above primers were blasted to retrieve sequence of FprA from genebank resulting in the following sequence:
+
<br>The above primers were blasted to retrieve sequence of FprA from genebank resulting in the following sequence:</p>
 +
<p style="font-family: courier">
 +
<br>tatgactcat atgcgtccct attacatcgc catcgtgggc tccgggccgt
 +
<br>cggcgttctt cgccgcggca tccttgctga aggccgccga cacgaccgag
 +
<br>gacctcgaca tggccgtcga catgctggag atgttgccga ctccctgggg
 +
<br>gctggtgcgc tccggggtcg cgccggatca ccccaagatc aagtcgatca
 +
<br>gcaagcaatt cgaaaagacg gccgaggacc cccgcttccg cttcttcggc
 +
<br>aatgtggtcg tcggcgaaca cgtccagccc ggcgagctct ccgagcgcta
 +
<br>cgacgccgtg atctacgccg tcggcgcgca gtccgatcgc atgttgaaca
 +
<br>tccccggtga ggacctgccg ggcagtatcg ccgccgtcga tttcgtcggc
 +
<br>tggtacaacg cacatccaca cttcgagcag gtatcacccg atctgtcggg
 +
<br>cgcccgggcc gtagttatcg gcaatggaaa cgtcgcgcta gacgtggcac
 +
<br>ggattctgct caccgatccc gacgtgttgg cacgcaccga tatcgccgat
 +
<br>cacgctttgg aatcgctacg cccacgcggt atccaggagg tggtgatcgt
 +
<br>cgggcgccga ggtccgctgc aggccgcgtt caccacgttg gagttgcgcg
 +
<br>agctggccga cctcgacggg gttgacgtgg tgatcgatcc ggcggagctg
 +
<br>gacggcatta ccgacgagga cgcggccgcg gtgggcaagg tctgcaagca
 +
<br>gaacatcaag gtgctgcgtg gctatgcgga ccgcgaaccc cgcccgggac
 +
<br>accgccgcat ggtgttccgg ttcttgacct ctccgatcga gatcaagggc
 +
<br>aagcgcaaag tggagcggat cgtgctgggc cgcaacgagc tggtctccga
 +
<br>cggcagcggg cgagtggcgg ccaaggacac cggcgagcgc gaggagctgc
 +
<br>cagctcagct ggtcgtgcgg tcggtcggct accgcggggt gcccacgccc
 +
<br>gggctgccgt tcgacgacca gagcgggacc atccccaacg tcggcggccg
 +
<br>aatcaacggc agccccaacg aatacgtcgt cgggtggatc aagcgcgggc
 +
<br>cgaccggggt gatcgggacc aacaagaagg acgcccaaga caccgtcgac
 +
<br>accttgatca agaatcttgg caacgccaag gagggcgccg agtgcaagag
 +
<br>ctttccggaa gatcatgccg accaggtggc cgactggcta gcagcacgcc
 +
<br>agccgaagct ggtcacgtcg gcccactggc aggtgatcga cgctttcgag
 +
<br>cgggccgccg gcgagccgca cgggcgtccc cgggtcaagt tggccagcct
 +
<br>ggccgagctg ttgcggattg ggctcggctg aggatcccgg at</p>
 +
<p>As previously, the sequence was imported into geneious, the biobrick restriction sites were manually removed from the sequence and restriction sites were added to the 5’ and 3’ ends in the same fashion as TB sirA.  However, in the case of FdrA, the restriction sites added for cloning into the vector pCOLAduet-1 were NdeI and AvrII.  As a result, there was no need to add an additional glycine after the start codon.  The resulting sequence is as follows:</p>
 +
<p style="font-family: courier">
 +
tatgaattct agatcatatg cgtccctatt acatcgccat cgtgggctcc
 +
<br>gggccgtcgg cgttcttcgc cgcggcatcc ttgctgaagg ccgccgacac
 +
<br>gaccgaggac ctcgacatgg ccgtcgacat gctggagatg ttgccgactc
 +
<br>cctgggggct ggtgcgctcc ggggtcgcgc cggatcaccc caagatcaag
 +
<br>tcgatcagca agcaattcga aaagacggcc gaggaccccc gcttccgctt
 +
<br>cttcggcaat gtggtcgtcg gcgaacacgt ccagcccggc gagctctccg
 +
<br>agcgctacga cgccgtgatc tacgccgtcg gcgcgcagtc cgatcgcatg
 +
<br>ttgaacatcc ccggtgagga cctgccgggc agtatcgccg ccgtcgattt
 +
<br>cgtcggctgg tacaacgcac atccacactt cgagcaggta tcacccgatc
 +
<br>tgtcgggcgc ccgggccgta gttatcggca atggaaacgt cgcgctagac
 +
<br>gtggcacgga ttctgctcac cgatcccgac gtgttggcac gcaccgatat
 +
<br>cgccgatcac gctttggaat cgctacgccc acgcggtatc caggaggtgg
 +
<br>tgatcgtcgg gcgccgaggt ccgcttcagg ccgcgttcac cacgttggag
 +
<br>ttgcgcgagc tggccgacct cgacggggtt gacgtggtga tcgatccggc
 +
<br>ggagctggac ggcattaccg acgaggacgc ggccgcggtg ggcaaggtct
 +
<br>gcaagcagaa catcaaggtg ctgcgtggct atgcggaccg cgaaccccgc
 +
<br>ccgggacacc gccgcatggt gttccggttc ttgacctctc cgatcgagat
 +
<br>caagggcaag cgcaaagtgg agcggatcgt gctgggccgc aacgagctgg
 +
<br>tctccgacgg cagcgggcga gtggcggcca aggacaccgg cgagcgcgag
 +
<br>gagctgccag ctcagctggt cgtgcggtcg gtcggctacc gcggggtgcc
 +
<br>cacgcccggg ctgccgttcg acgaccagag cgggaccatc cccaacgtcg
 +
<br>gcggccgaat caacggcagc cccaacgaat acgtcgtcgg gtggatcaag
 +
<br>cgcgggccga ccggggtgat cgggaccaac aagaaggacg cccaagacac
 +
<br>cgtcgacacc ttgatcaaga atcttggcaa cgccaaggag ggcgccgagt
 +
<br>gcaagagctt tccggaagat catgccgacc aggtggccga ctggctagca
 +
<br>gcacgccagc cgaagctggt cacgtcggcc cactggcagg tgatcgacgc
 +
<br>tttcgagcgg gccgccggcg agccgcacgg gcgtccccgg gtcaagttgg
 +
<br>ccagcctggc cgagctgttg cggattgggc tcggctgagg atccctagga
 +
<br>ctagtctgca gttacg</p>
 +
<p><em>Mycobacterium smegmatis</em>
 +
<br>We didn't find a reference of M. Smegmatis FprA being cloned into E. coli so we used the following sequenced data:</p>
 +
<p style="font-family: courier">
 +
<br>atgcgcccgt accacgtagc gatcgtcggc tcaggaccct ccggattctt
 +
<br>tgctgccgca tcgctgctga agtttgccga ctctcaaccc gaccgcgatg
 +
<br>tccgcgtgga catgctcgag atgctgccga ccccgtgggg tctggtgcgc
 +
<br>tccggcgtcg ctcccgacca tccgaagatc aagtcgatca gcgcccagtt
 +
<br>cgagaagacc gcggccgacc cccgcttccg gttcttcggc aacgtccggg
 +
<br>tcggtgagca cgtccaaccg ggcgaactcg ccgaacgcta cgacgccgtc
 +
<br>gtctacgcca ccggcgcaca gtccgaccgc gcgctgaaca tccccggcga
 +
<br>ggagctgccg ggcagcgtcg cggccgtcga cttcgtgggt tggtacaacg
 +
<br>cgcatccgca tttccgcgag atggcgcccg atctctccgg cgggcgcgcc
 +
<br>gtggtggtcg gcaacggcaa cgtcgcactc gatgtcgcac gcatcctggt
 +
<br>cagcgatccg aaggcgttgg ccaacaccga cattgccgac catgcgctgg
 +
<br>acaagctgga cacgcgcggt gtggacgagg tcgtggtgct cggccggcgt
 +
<br>ggtccgctgc aggcgacgtt caccacgctg gagttgcgtg aactcggcga
 +
<br>catggagggc ctcggcgacg tcgacgtgat cctggatccg gccgatttcg
 +
<br>ccgacatcac cgacgaggat ctcgaagccg cgggcaagac cgtcaagcag
 +
<br>aacatcaagg tgctgcgcgg ttacgccgag cgggaaccgc gaggcgccaa
 +
<br>gcggcgcatc gtgttccggt tcttcacctc accgatcgag ctgcgcggcg
 +
<br>aggaccgcgt ggaatcgatc gtgttgggac gcaacgaact cgtgcgcggc
 +
<br>gccgacggca ggatggtggc caaggacacc ggtgagcgcg aagaactgcc
 +
<br>ggcacagctg gtggtgcgtg cggtcggata ccgcggcgtg ccgactccag
 +
<br>gcctgccgtt cgacgagcgt tcgggcacca tcccgcacac cgacggccgc
 +
<br>gtcgagggca gcgccaacga gtacgtggtg ggctggatca agcgcggacc
 +
<br>gtccggcgtc atcggcagca acaagaagga ctcacaggac accgtcaaca
 +
<br>ccctgctcga cgacctcgcc gcggcccggc tgccccagcg cgggcccgac
 +
<br>cactccgaga agctcgcggc gtggttgctg gaacgtcagc ccaaggtggt
 +
<br>cacgggcgag cactggaagc tgatcgacga ctacgagcgc gccgcgggcg
 +
<br>aaccgaccgg ccgaccccgg gtgaagctgg cgagcgtggc tgaactgctg
 +
<br>cgcgtgggcc acggctga</p>
 +
<p>As above, the sequence was imported into geneious, the biobrick restriction sites were manually removed from the sequence and restriction sites were added to the 5’ and 3’ ends in the same fashion as TB sirA.  Again, the restriction sites added for cloning into the vector pCOLAduet-1 were NdeI and AvrII.  As a result, there was no need to add an additional glycine after the start codon.  The resulting sequence is as follows:</p>
 +
<p style="font-family: courier">
 +
<br>tatgaattct agatcatatg cgcccgtacc acgtagcgat cgtcggctca
 +
<br>ggaccctccg gattctttgc tgccgcatcg ctgctgaagt ttgccgactc
 +
<br>tcaacccgac cgcgatgtcc gcgtggacat gctcgagatg ctgccgaccc
 +
<br>cgtggggtct ggtgcgctcc ggcgtcgctc ccgaccatcc gaagatcaag
 +
<br>tcgatcagcg cccagttcga gaagaccgcg gccgaccccc gcttccggtt
 +
<br>cttcggcaac gtccgggtcg gtgagcacgt ccaaccgggc gaactcgccg
 +
<br>aacgctacga cgccgtcgtc tacgccaccg gcgcacagtc cgaccgcgcg
 +
<br>ctgaacatcc ccggcgagga gctgccgggc agcgtcgcgg ccgtcgactt
 +
<br>cgtgggttgg tacaacgcgc atccgcattt ccgcgagatg gcgcccgatc
 +
<br>tctccggcgg gcgcgccgtg gtggtcggca acggcaacgt cgcactcgat
 +
<br>gtcgcacgca tcctggtcag cgatccgaag gcgttggcca acaccgacat
 +
<br>tgccgaccat gcgctggaca agctggacac gcgcggtgtg gacgaggtcg
 +
<br>tggtgctcgg ccggcgtggt ccgcttcagg cgacgttcac cacgctggag
 +
<br>ttgcgtgaac tcggcgacat ggagggcctc ggcgacgtcg acgtgatcct
 +
<br>ggatccggcc gatttcgccg acatcaccga cgaggatctc gaagccgcgg
 +
<br>gcaagaccgt caagcagaac atcaaggtgc tgcgcggtta cgccgagcgg
 +
<br>gaaccgcgag gcgccaagcg gcgcatcgtg ttccggttct tcacctcacc
 +
<br>gatcgagctg cgcggcgagg accgcgtgga atcgatcgtg ttgggacgca
 +
<br>acgaactcgt gcgcggcgcc gacggcagga tggtggccaa ggacaccggt
 +
<br>gagcgcgaag aactgccggc acagctggtg gtgcgtgcgg tcggataccg
 +
<br>cggcgtgccg actccaggcc tgccgttcga cgagcgttcg ggcaccatcc
 +
<br>cgcacaccga cggccgcgtc gagggcagcg ccaacgagta cgtggtgggc
 +
<br>tggatcaagc gcggaccgtc cggcgtcatc ggcagcaaca agaaggactc
 +
<br>acaggacacc gtcaacaccc tgctcgacga cctcgccgcg gcccggctgc
 +
<br>cccagcgcgg gcccgaccac tccgagaagc tcgcggcgtg gttgctggaa
 +
<br>cgtcagccca aggtggtcac gggcgagcac tggaagctga tcgacgacta
 +
<br>cgagcgcgcc gcgggcgaac cgaccggccg accccgggtg aagctggcga
 +
<br>gcgtggctga actgctgcgc gtgggccacg gctgacctag gactagtctg
 +
<br>cagttacg
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Latest revision as of 11:39, 12 August 2013

Drug Screening

Tuesday 11th June

Design of FdrA genes for synthesis

FdrA

Mycobacterium tuberculosis
Reference: McLean et al. 2003. Kinetic, spectroscopic and thermodynamic characterization of the Mycobacterium tuberculosis adrenodoxin reductase homologue FprA. doi: 10.1042/BJ20021692
Primers:
MtbfdrA F      TATGACTCATATGCGTCCCTATTACATCGCCATCG
MtbfdrA R      ATCCGGGATCCTCAGCCGAGCCCAATCCGCAACAGC
The above primers were blasted to retrieve sequence of FprA from genebank resulting in the following sequence:


tatgactcat atgcgtccct attacatcgc catcgtgggc tccgggccgt
cggcgttctt cgccgcggca tccttgctga aggccgccga cacgaccgag
gacctcgaca tggccgtcga catgctggag atgttgccga ctccctgggg
gctggtgcgc tccggggtcg cgccggatca ccccaagatc aagtcgatca
gcaagcaatt cgaaaagacg gccgaggacc cccgcttccg cttcttcggc
aatgtggtcg tcggcgaaca cgtccagccc ggcgagctct ccgagcgcta
cgacgccgtg atctacgccg tcggcgcgca gtccgatcgc atgttgaaca
tccccggtga ggacctgccg ggcagtatcg ccgccgtcga tttcgtcggc
tggtacaacg cacatccaca cttcgagcag gtatcacccg atctgtcggg
cgcccgggcc gtagttatcg gcaatggaaa cgtcgcgcta gacgtggcac
ggattctgct caccgatccc gacgtgttgg cacgcaccga tatcgccgat
cacgctttgg aatcgctacg cccacgcggt atccaggagg tggtgatcgt
cgggcgccga ggtccgctgc aggccgcgtt caccacgttg gagttgcgcg
agctggccga cctcgacggg gttgacgtgg tgatcgatcc ggcggagctg
gacggcatta ccgacgagga cgcggccgcg gtgggcaagg tctgcaagca
gaacatcaag gtgctgcgtg gctatgcgga ccgcgaaccc cgcccgggac
accgccgcat ggtgttccgg ttcttgacct ctccgatcga gatcaagggc
aagcgcaaag tggagcggat cgtgctgggc cgcaacgagc tggtctccga
cggcagcggg cgagtggcgg ccaaggacac cggcgagcgc gaggagctgc
cagctcagct ggtcgtgcgg tcggtcggct accgcggggt gcccacgccc
gggctgccgt tcgacgacca gagcgggacc atccccaacg tcggcggccg
aatcaacggc agccccaacg aatacgtcgt cgggtggatc aagcgcgggc
cgaccggggt gatcgggacc aacaagaagg acgcccaaga caccgtcgac
accttgatca agaatcttgg caacgccaag gagggcgccg agtgcaagag
ctttccggaa gatcatgccg accaggtggc cgactggcta gcagcacgcc
agccgaagct ggtcacgtcg gcccactggc aggtgatcga cgctttcgag
cgggccgccg gcgagccgca cgggcgtccc cgggtcaagt tggccagcct
ggccgagctg ttgcggattg ggctcggctg aggatcccgg at

As previously, the sequence was imported into geneious, the biobrick restriction sites were manually removed from the sequence and restriction sites were added to the 5’ and 3’ ends in the same fashion as TB sirA. However, in the case of FdrA, the restriction sites added for cloning into the vector pCOLAduet-1 were NdeI and AvrII. As a result, there was no need to add an additional glycine after the start codon. The resulting sequence is as follows:

tatgaattct agatcatatg cgtccctatt acatcgccat cgtgggctcc
gggccgtcgg cgttcttcgc cgcggcatcc ttgctgaagg ccgccgacac
gaccgaggac ctcgacatgg ccgtcgacat gctggagatg ttgccgactc
cctgggggct ggtgcgctcc ggggtcgcgc cggatcaccc caagatcaag
tcgatcagca agcaattcga aaagacggcc gaggaccccc gcttccgctt
cttcggcaat gtggtcgtcg gcgaacacgt ccagcccggc gagctctccg
agcgctacga cgccgtgatc tacgccgtcg gcgcgcagtc cgatcgcatg
ttgaacatcc ccggtgagga cctgccgggc agtatcgccg ccgtcgattt
cgtcggctgg tacaacgcac atccacactt cgagcaggta tcacccgatc
tgtcgggcgc ccgggccgta gttatcggca atggaaacgt cgcgctagac
gtggcacgga ttctgctcac cgatcccgac gtgttggcac gcaccgatat
cgccgatcac gctttggaat cgctacgccc acgcggtatc caggaggtgg
tgatcgtcgg gcgccgaggt ccgcttcagg ccgcgttcac cacgttggag
ttgcgcgagc tggccgacct cgacggggtt gacgtggtga tcgatccggc
ggagctggac ggcattaccg acgaggacgc ggccgcggtg ggcaaggtct
gcaagcagaa catcaaggtg ctgcgtggct atgcggaccg cgaaccccgc
ccgggacacc gccgcatggt gttccggttc ttgacctctc cgatcgagat
caagggcaag cgcaaagtgg agcggatcgt gctgggccgc aacgagctgg
tctccgacgg cagcgggcga gtggcggcca aggacaccgg cgagcgcgag
gagctgccag ctcagctggt cgtgcggtcg gtcggctacc gcggggtgcc
cacgcccggg ctgccgttcg acgaccagag cgggaccatc cccaacgtcg
gcggccgaat caacggcagc cccaacgaat acgtcgtcgg gtggatcaag
cgcgggccga ccggggtgat cgggaccaac aagaaggacg cccaagacac
cgtcgacacc ttgatcaaga atcttggcaa cgccaaggag ggcgccgagt
gcaagagctt tccggaagat catgccgacc aggtggccga ctggctagca
gcacgccagc cgaagctggt cacgtcggcc cactggcagg tgatcgacgc
tttcgagcgg gccgccggcg agccgcacgg gcgtccccgg gtcaagttgg
ccagcctggc cgagctgttg cggattgggc tcggctgagg atccctagga
ctagtctgca gttacg

Mycobacterium smegmatis
We didn't find a reference of M. Smegmatis FprA being cloned into E. coli so we used the following sequenced data:


atgcgcccgt accacgtagc gatcgtcggc tcaggaccct ccggattctt
tgctgccgca tcgctgctga agtttgccga ctctcaaccc gaccgcgatg
tccgcgtgga catgctcgag atgctgccga ccccgtgggg tctggtgcgc
tccggcgtcg ctcccgacca tccgaagatc aagtcgatca gcgcccagtt
cgagaagacc gcggccgacc cccgcttccg gttcttcggc aacgtccggg
tcggtgagca cgtccaaccg ggcgaactcg ccgaacgcta cgacgccgtc
gtctacgcca ccggcgcaca gtccgaccgc gcgctgaaca tccccggcga
ggagctgccg ggcagcgtcg cggccgtcga cttcgtgggt tggtacaacg
cgcatccgca tttccgcgag atggcgcccg atctctccgg cgggcgcgcc
gtggtggtcg gcaacggcaa cgtcgcactc gatgtcgcac gcatcctggt
cagcgatccg aaggcgttgg ccaacaccga cattgccgac catgcgctgg
acaagctgga cacgcgcggt gtggacgagg tcgtggtgct cggccggcgt
ggtccgctgc aggcgacgtt caccacgctg gagttgcgtg aactcggcga
catggagggc ctcggcgacg tcgacgtgat cctggatccg gccgatttcg
ccgacatcac cgacgaggat ctcgaagccg cgggcaagac cgtcaagcag
aacatcaagg tgctgcgcgg ttacgccgag cgggaaccgc gaggcgccaa
gcggcgcatc gtgttccggt tcttcacctc accgatcgag ctgcgcggcg
aggaccgcgt ggaatcgatc gtgttgggac gcaacgaact cgtgcgcggc
gccgacggca ggatggtggc caaggacacc ggtgagcgcg aagaactgcc
ggcacagctg gtggtgcgtg cggtcggata ccgcggcgtg ccgactccag
gcctgccgtt cgacgagcgt tcgggcacca tcccgcacac cgacggccgc
gtcgagggca gcgccaacga gtacgtggtg ggctggatca agcgcggacc
gtccggcgtc atcggcagca acaagaagga ctcacaggac accgtcaaca
ccctgctcga cgacctcgcc gcggcccggc tgccccagcg cgggcccgac
cactccgaga agctcgcggc gtggttgctg gaacgtcagc ccaaggtggt
cacgggcgag cactggaagc tgatcgacga ctacgagcgc gccgcgggcg
aaccgaccgg ccgaccccgg gtgaagctgg cgagcgtggc tgaactgctg
cgcgtgggcc acggctga

As above, the sequence was imported into geneious, the biobrick restriction sites were manually removed from the sequence and restriction sites were added to the 5’ and 3’ ends in the same fashion as TB sirA. Again, the restriction sites added for cloning into the vector pCOLAduet-1 were NdeI and AvrII. As a result, there was no need to add an additional glycine after the start codon. The resulting sequence is as follows:


tatgaattct agatcatatg cgcccgtacc acgtagcgat cgtcggctca
ggaccctccg gattctttgc tgccgcatcg ctgctgaagt ttgccgactc
tcaacccgac cgcgatgtcc gcgtggacat gctcgagatg ctgccgaccc
cgtggggtct ggtgcgctcc ggcgtcgctc ccgaccatcc gaagatcaag
tcgatcagcg cccagttcga gaagaccgcg gccgaccccc gcttccggtt
cttcggcaac gtccgggtcg gtgagcacgt ccaaccgggc gaactcgccg
aacgctacga cgccgtcgtc tacgccaccg gcgcacagtc cgaccgcgcg
ctgaacatcc ccggcgagga gctgccgggc agcgtcgcgg ccgtcgactt
cgtgggttgg tacaacgcgc atccgcattt ccgcgagatg gcgcccgatc
tctccggcgg gcgcgccgtg gtggtcggca acggcaacgt cgcactcgat
gtcgcacgca tcctggtcag cgatccgaag gcgttggcca acaccgacat
tgccgaccat gcgctggaca agctggacac gcgcggtgtg gacgaggtcg
tggtgctcgg ccggcgtggt ccgcttcagg cgacgttcac cacgctggag
ttgcgtgaac tcggcgacat ggagggcctc ggcgacgtcg acgtgatcct
ggatccggcc gatttcgccg acatcaccga cgaggatctc gaagccgcgg
gcaagaccgt caagcagaac atcaaggtgc tgcgcggtta cgccgagcgg
gaaccgcgag gcgccaagcg gcgcatcgtg ttccggttct tcacctcacc
gatcgagctg cgcggcgagg accgcgtgga atcgatcgtg ttgggacgca
acgaactcgt gcgcggcgcc gacggcagga tggtggccaa ggacaccggt
gagcgcgaag aactgccggc acagctggtg gtgcgtgcgg tcggataccg
cggcgtgccg actccaggcc tgccgttcga cgagcgttcg ggcaccatcc
cgcacaccga cggccgcgtc gagggcagcg ccaacgagta cgtggtgggc
tggatcaagc gcggaccgtc cggcgtcatc ggcagcaaca agaaggactc
acaggacacc gtcaacaccc tgctcgacga cctcgccgcg gcccggctgc
cccagcgcgg gcccgaccac tccgagaagc tcgcggcgtg gttgctggaa
cgtcagccca aggtggtcac gggcgagcac tggaagctga tcgacgacta
cgagcgcgcc gcgggcgaac cgaccggccg accccgggtg aagctggcga
gcgtggctga actgctgcgc gtgggccacg gctgacctag gactagtctg
cagttacg